In Vivo Functional Real-Time Imaging of Transplanted Islets Using Positron Emission Tomography (PET) Positron emission tomography (PET) is a established quantitative and noninvasive imaging modality. With the PET reporter gene (PRG)/PET reporter probe (PRP) system, based on a mutant form of herpes simplex virus 1 thymidine kinase (HSV1-sr39tk), the PET signal is directly proportional to the enzymatic activity of sr39TK9-14. In this protocol, we describe in detail a method for reporter gene labeling of islets and quantitative scanning using a reporter probe.
Live Imaging of Caenorhabditis elegans: Preparation of Samples Caenorhabditis elegans, a small (adults are ~1 mm long), free-living soil nematode that feeds on bacteria, is an ideal organism for applying various live microscopy techniques. This protocol describes useful techniques for preparing C. elegans for live microscopic analysis. Details of sample preparation depend on the developmental stage of the worm to be studied.
Protocol Live Imaging of Caenorhabditis Elegans To image early cleavages and chromatin dynamics, it is convenient to use histone H2B fused to GFP or lamin::GFP. Time-lapse movies can be obtained using conventional confocal microscope systems and their included software. Early embryos dissected from transgenic hermaphrodites are placed with egg salts on agar pads. Chromatin dynamics can be followed easily, and wild-type embryonic cells can be compared with mutants or RNAi-treated embryos.
Selection of Appropriate Imaging Equipment and Methodology for Live Cell Imaging in Drosophila In the past two decades, there have been many revolutions in light microscopy techniques made possible by improvements in optics, detector technology, and computers. Furthermore, there is no indication that the rate of development of new equipment is slowing down. Here we attempt to provide an overview of available options and important considerations applicable to imaging Drosophila cells and tissues.
Time-Lapse Cinematography in Living Drosophila Tissues: Preparation of Material Sophisticated fluorescence microscopy methods & equipment, now allow cellular events to be studied at high resolution in living material. The studying of living fly tissues presents unique difficulties in keeping the cells alive, introducing fluorescent probes, & imaging through thick hazy cytoplasm. This protocol outlines the preparation of major tissue types amenable to study by time-lapse cinematography and different methods for keeping them alive.